文章摘要
高关刚,盛园园,张嘉杰,刘佳明,徐 俐.Bodipy-C12在癌细胞过氧化物酶体中富集[J].,2020,(1):1-7
Bodipy-C12在癌细胞过氧化物酶体中富集
Bodipy-C12 can be Enriched in Peroxisomes of Cancer Cells
投稿时间:2019-07-11  修订日期:2019-08-08
DOI:10.13241/j.cnki.pmb.2020.01.001
中文关键词: 脂肪酸  线粒体  脂滴  过氧化物酶体  Bodipy-C12
英文关键词: Fatty acid  Mitochondria  Lipid droplet  Peroxisomes  Bodipy-C12
基金项目:
作者单位E-mail
高关刚 清华大学生命科学学院 北京 100084 ggg12@mails.tsinghua.edu.cn 
盛园园 清华大学生命科学学院 北京 100084  
张嘉杰 清华大学生命科学学院 北京 100084  
刘佳明 清华大学生命科学学院 北京 100084  
徐 俐 清华大学生命科学学院 北京 100084  
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中文摘要:
      摘要 目的:研究并比较Bodipy标记的月桂酸(Bodipy-C12)在癌细胞与正常细胞中的亚细胞定位。方法:在多种癌细胞和正常细胞的培养基中加入Bodipy-C12(1 μg/mL),利用共聚焦显微镜活细胞定时间隔拍摄,结合不同细胞器的分子标记蛋白,观察Bodipy-C12五分钟内在不同细胞器中的定位。结果:在人肝癌细胞系HepG2细胞中,Bodpy-C12信号不仅仅存在于线粒体和脂滴,同时富集于过氧化物酶体中。我们分别采用Pex3-GFP、Pex14-GFP、GFP-Pex16、GFP-SKL和GFP-Pmp34等特异性定位的过氧化物酶体蛋白,确认Bodpy-C12信号富集于过氧化物酶体。此外,过氧化物酶体中Bodpy-C12信号的富集发生在更多的癌细胞系中,例如结肠癌细胞HCT116和乳腺癌细胞MCF7。不同的是,在正常细胞系如3T3-L1,NRK和COS7中,Bodipy-C12信号存在于线粒体和脂滴中,但未在过氧化物酶体中检测到。结论:Bodipy-C12信号存在于正常细胞和癌细胞的脂滴和线粒体中,且其在癌细胞过氧化物酶体中富集,而不存在于正常细胞的过氧化物酶体中,预示癌细胞中过氧化物酶体脂代谢的差异。
英文摘要:
      ABSTRACT Objective: To investigate and compare the subcellular localization of Bodipy-labeled lauric acid (Bodipy-C12) in cancer cells and normal cells. Methods: Various cancer cells and normal cells were cultured in medium with Bodipy-C12 (1 μg/mL). We detected the signal of Bodipy-C12 in different organelles labeled by specific molecular maker proteins with using live-cell time-lapse confocal microscope in 5 minutes. Results: In a human hepatocellular carcinoma cell line HepG2, the signal of Bodpy-C12 was not only detected in lipid droplet and mitochondria, but it can also be enriched in peroxisomes. The co-localization of Bodpy-C12 signal with specific molecular protein markers, including Pex3-GFP, Pex14-GFP, GFP-Pex16, GFP-SKL and GFP-Pmp34, verified the existence of Bodpy-C12 in peroxisomes. Further, the enrichment of Bodpy-C12 in peroxisomes occured to more cancer cell lines such as colorectal cancer cell HCT116 and breast cancer cell MCF7. Differently, Bodpy-C12 signal was detected only in lipid droplet and mitochondria, but not in peroxisomes of normal cell lines such as 3T3-L1, NRK, COS7. Conclusion: Bodipy-C12 signal existed in lipid droplet and mitochondria of normal cells and cancer cells, and it can be enriched in peroxisomes of cancer cells, but not in peroxisomes of normal cells, indicating a different lipid metabolism of the peroxisomes in cancer cells.
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